Methods for regulating potency of pluripotent stem cells and applications thereof

a technology of pluripotent stem cells and potency regulation, applied in the field of pluripotent stem cell potency regulation, can solve the problems of rising immunorejection problems, hpscs' importance of cholesterol, and insufficient study of transmembrane proteins, etc., to achieve the effect of boosting the reprogramming efficiency of somatic cells

Pending Publication Date: 2022-03-24
ACAD SINIC
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  • Abstract
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Benefits of technology

[0031]Further, it is found in the present invention that cholesterol can boost the reprogramming efficiency of somatic cells, such as skin cells e.g. fibroblast. Therefore, the present invention provides a method for generating pluripotent stem cells (iPSCs) comprising culturing somatic cells in a condition which allows a proportion of the skin cells to dedifferentiate into iPSCs, wherein the condition comprises a culture medium which comprises cholesterol. In some embodiments, the somatic cells are genetic engineered, for example, by introduced with a recombinant nucleic acid, to overexpress one or more reprograming factors, for example, OSKM including Oct4, Sox2, Klf4, and cMyc. Also provided is use of cholesterol for treating somatic cells for generating pluripotent stem cells (iPSCs) therefrom via reprograming. Further provided is a composition e.g. medium composition comprising cholesterol and a basic medium which is useful in treating somatic cells for generating pluripotent stem cells (iPSCs) therefrom via reprograming.

Problems solved by technology

The potential of ESCs in regenerative medicine is enormous, but it rises immunorejection problems.
However, the transmembrane proteins have not been studied in detail.
However, the importance of cholesterol in human pluripotent stem cell (hPSCs) remain elusive.

Method used

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  • Methods for regulating potency of pluripotent stem cells and applications thereof
  • Methods for regulating potency of pluripotent stem cells and applications thereof
  • Methods for regulating potency of pluripotent stem cells and applications thereof

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[0075]Except for the well characterized function in tumor metastasis, transmembrane glycoprotein podocalyxin-like protein 1 (PODXL, also named as podocalyxin like protein-1, PCLP1, MEP21, Gp200 / GCTM-2, and Thrombomucin), function in hPSCs is not known. Here, we demonstrate the knockdown of PODXL in undifferentiated hPSCs significantly inhibited the self-renewal abilities that currently block the expression of c-MYC and telomerase proteins. Of note, the induction or reprogramming of induced pluripotent stem cells (iPSCs) and extended pluripotent stem cells (EPSCs) were severely blocked upon the knockdown of PODXL. Consistently, upregulation of PODXL facilitated hPSC renewal, enhance the expressions of c-MYC and telomerase, and promote iPSC / EPSC formation. In a microarray analysis, overexpression of PODXL activate HMGCR expression, which control the cholesterol biosynthesis. We found that PODXL also upregulates SREBP1 / 2 expression. Of note, hPSCs are more sensitive to cholesterol inhi...

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Abstract

The present invention relates to a method for regulating potency of pluripotent stem cells (PSCs) by modulating expression of podocalyxin-like protein 1 (PODXL) and applications thereof.

Description

RELATED APPLICATIONS[0001]This application claims the benefit of U.S. provisional application No. 62 / 784,942, filed Dec. 26, 2018 under 35 U.S.C. § 119, the entire content of which is incorporated herein by reference.TECHNOLOGY FIELD[0002]The present invention relates to a method for regulating potency of pluripotent stem cells (PSCs) by modulating expression of podocalyxin-like protein 1 (PODXL) and cholesterol and applications thereof.BACKGROUND OF THE INVENTION[0003]Human embryonic stem cells (hESCs), generated from the inner cell mass of the early embryo have the ability to unlimited proliferation and differentiate into endoderm, mesoderm, and ectoderm, and potentially into all cell types except placenta (Thomson et al., 1998). hESCs behave like epiblast cells and claimed as the primed state (Brons et al., 2007; Kumari, 2016; Nichols and Smith, 2009; Tesar et al., 2007). Switching the culture medium can change the primed state ESCs into naive-like state. Naïve stem cells are les...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/074C12N5/00C12N15/86C12N15/873C12N15/85C07K14/435
CPCC12N5/0696C12N5/0081C12N15/86C12N15/873C12N15/8509C12N2506/03C12N2740/15043C12N2506/1307C12N2506/02C12N2506/45C07K14/435C12N15/63C12N15/64C12N15/1138C12N2510/00C12N5/0606C12N2500/92C12N2501/235C12N2501/727C12N2533/90C12N2500/35C12N2310/14C12N2310/531
Inventor LU, JOYCE JEANCHEN, WEI-JU
Owner ACAD SINIC
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